Antigenic profile of human melanoma cells. Analysis with monoclonal antibodies to histocompatibility antigens and to melanoma-associated antigens (original) (raw)
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The Journal of experimental medicine, 1983
This study represents an initial attempt to analyze the humoral immune reactions of patients with malignant melanoma by hybridoma methodology. Using lymphocytes from regional lymph nodes, peripheral blood and tumor infiltrates, 158 fusions were performed with SKO-007 (human myeloma line), LICR-LON-HMy2 (LICR-2), GM 4672 (human lymphoblastoid lines), or NS-1 (mouse myeloma line). Fusion of lymph node lymphocytes with NS-1 resulted in a 3-4 times higher frequency of clones than fusion with LICR-2, and a 10 times higher frequency than fusion with SKO-007 or GM 4672. In the case of peripheral blood lymphocytes, fusion with NS-1 gave greater than 25 times higher frequency of clones than fusion with LICR-2 or SKO-007. Production of human mu, gamma, or alpha heavy chains was detected in 50-80% of wells containing growing clones, and the levels of immunoglobulin ranged from 0.3 micrograms to 40 micrograms/ml. NS-1-derived clones could be easily subcultured, while LICR-2 and SKO-007 clones g...
Common human melanoma-associated antigen(s) detected by monoclonal antibodies
Cancer research, 1980
Hybridoma cells have been derived from a fusion between mouse myeloma cells (P3-NSI/1Ag4) and spleen cells from a mouse immunized with membrane-enriched fractions from the human melanoma cell line Me-43. Of the 26 hybrids obtained, seven secreted antibodies which reacted with the melanoma cell line used for immunoassay. The specificity of the antibodies produced by the seven positive hybrids was further investigated on 16 melanoma cell lines, 15 other tumors, and 14 lymphoblastoid cell lines. The antibodies from four positive hybrids showed a broad reactivity, whereas those from three hybrids reacted exclusively with melanoma cells. The antibodies from two of these three hybrids, alpha-Mel/5 and alpha-Mel/14, seem to be directed against common melanoma antigen(s) since they reacted with all (with one exception) of the 16 melanoma cell lines tested only with five of the 16 melanoma lines. Reciprocal binding inhibition tests using [3H]leeucine-labeled antibodies showed that alpha-Mel/...
Cell surface antigens of human melanoma identified by monoclonal antibody
Proceedings of the National Academy of Sciences, 1979
Mouse NS-1 myeloma cells were fused with spleen cells from mice that had been immunized with cells from a human melanoma, M1804. Hybrid cells were grown in selective medium and tested for production of antibody to surface antigens of M1804 cells. Three hybrids that produced antibodies that bound to the melanoma cells but not to autologous skin fibroblasts were cloned. Antibodies produced by two of the clones were cytotoxic to M1804 cells in the presence of rabbit complement. Extensive specificity tests showed that the antibodies produced by the clones bound strongly only to M1804 cells; significant, although weaker, binding occurred with 2 of 11 allogeneic melanomas. Apart from weak binding of the antibody produced by one of the clones to a breast carcinoma, binding assays of five carcinomas, one sarcoma, and fibroblasts from 17 individuals were negative, as were cytotoxic tests of 10
Analysis of two human monoclonal antibodies against melanoma
International Journal of Cancer, 1992
B cells derived from peripheral-blood lymphocytes (PBL) and tumor-infiltrating lymphocytes (TlL) from a patient with a high serum antibody titer to autologous melanoma were transformed with Epstein-Barr virus (EBV) and evaluated for reactivity against autologous tumor. B cells producing antibody reactive with autologous tumor and unreactive with normal fibroblasts were detected both in TIL and in PBL. One cell line derived from PBL and another derived from TIL sustained production of tumor-reactive antibody for I 0 weeks and over I 5 months respectively. The cell line derived from PBL, 2DI I, produced an antibody reactive with a tryprin-resistant antigen expressed on the cell membrane of autologous and allogeneic melanoma cell lines. The cell line derived from TIL, IF6, produced an antibody reactive with a cell-surface glycoprotein expressed by 5 autologous melanoma cell lines derived from 5 different metastases and I6/ I 9 allogeneic melanoma cell lines. IF6 also showed reactivity with cell lines derived from a blue nevus, a congenital nevus, an astrocytoma, and I /4 renal-cell carcinomas; but it was not reactive with 5 foreskin melanocyte cell lines, 2 normal fibroblast lines, 5 leukemia/lymphoma lines, 8 lung-cancer lines, 8 glioblastoma lines, or lines derived from I ovarian carcinoma, I colon carcinoma, I vulvar carcinoma, I fibrosarcoma, I murine melanoma, or 4 murine leukemia/ lymphomas. We describe here an antibody that detects a new melanoma specificity obtained by EBV transformation of tumorinfiltrating B cells.
International Journal of Cancer, 1976
Antibody-dependent cell-mediated cytotoxic assays have been used to examine antigens on human melanoma cells obtained either directly from patients or from long-term melanoma cell lines. A panel of melanoma antisera was selected from human subjects which could be shown not to have significant reactivity to histocompatibility antigens. With these antisera extensive cross-reactions between melanoma cells were found. However, the cross-reactivity was incomplete and the pattern of reactivity was dijerent for each antiserum tested. These results were not consistent with a common melanoma antigen on human melanoma cells but rather indicated heterogeneity of melanoma antigens and multiple antibody specificities in the sera tested. This appeared to be confirmed by extensive cross-absorption studies which indicated limited crossreactivity of antigens on melanoma cells from either long-term or short-term cultures. Several changes in the antigenic profile of melanoma cells in vitro from both long-term and short-term cultures were documented which resulted from contamination of the melanoma cell lines with non-melanoma cells and fibroblasts. Melanoma antisera tnay therefore be useful to monitor changes in long-term cultures which would otherwise give spurious results in in vitro tests. These results appear to have considerable significance for understanding tumourjhost relationships and for the establishment of rational imrnunotherapeutic procedures and diagnostic tests in melanoma.
Cancer research, 1979
To characterize the antigen present on the surface of cultured human malignant melanoma, three monkey xeno geneic antisera were raised. After appropriate absorption with pooled human erythrocytes, peripheral blood leuko cytes, and liven homogenate, no blood group or HLA neac tivity was detectable. Analysis of melanoma antigenic spec ificity was performed by the mixed hemadsonption microas say on live monolayer cells in conjunction with quantitative microabsorption analysis. To eliminate reactivity against nonmelanoma lines, 2 of the 3 antisera required further absorption with cells of the KB oral carcinoma line. The absorbed antisera reacted with 9 of 10 long-term estab lished lines, 3 of 3 short-term cultures of human malignant melanoma, and i of 10 nonmelanoma epithelial and fibro blastic cell lines. Absorption experiments using a variety of cultured cells and fresh tissue homogenates of adult human melanoma and nonmelanoma sources further substantiated the results obtained with the direct tests. While each mela noma line showed a differing degree and pattern of neactiv ity, the antisera were most reactive against their respective immunizing lines as revealed by both the direct tests and absorption analysis. By quantitative absorption analysis, no evidence for individually specific melanoma antigens was obtained. The only positive absorption with nonmelanoma adult tissues was obtained with a retinoblastoma cell line, indicating the presence of antigens shared by tumors of common neunoectodermal origin. Extensive absorption with two xenogeneic malignant melanoma (munine and porcine) homogenates, normal human adult skin, and spleen tissues, or Bacillus Calmette-Guéninfailed to reduce the reactivity against melanoma-associated antigens. Fur then absorption with human fetal tissues of 8 to 20 weeks of gestation removed part but not all of the reactivity. These studies with xenogeneic monkey antisera provide evidence for the existence of common melanoma-associated anti gens as well as distinct but shared human fetal antigens on human melanoma cells. , This is Paper 8 of the series, â€oeCharacterization of Human Malignant MelanomaCell Lines.― The work was supportedby the Medical Research Council of Canada and the Ontario Cancer Treatment and Research Foun dation.
Human melanoma-specific oncofetal antigen defined by a mouse monoclonal …
… Journal of Cancer
140.240, an lgC2a mouse monoclonal antibody raised against a cultured human melanoma cell line, was highly specific for melanoma cells as determined by direct and absorption analyses in a mixed hemadsorption assay. Supematants of doubly cloned hybridomas producing antibody 140.240 reacted with all cultured and fresh melanomas tested but failed to react with a variety of carcinomas, sarcomas, lymphomas, leukemias and other tumors of neuroectoderrnal origin. This antibody did not react with B-lymphoid cell lines, ruling out HLA-DR specificity. Non-reactivity of antibody 140.240 with peripheral blood lymphocytes obtained from the donor of the immunizing melanoma line excluded the possibility of detecting histocompatibility antigens. Nevus cells were also non-reactive. However, antibody 140.240 did identify an antigenic determinant on tissue homogenates prepared from fetuses of 10-14 weeks' gestation. The antigen involved was shed by cultured melanoma lines and, by immunoprecipitation analysis of radiolabelled lysates, had a molecular weight of approximately 87kdal. Thus, the structure identified by monoclonal antibody 140.240 is a melanoma-specific oncofetal antigen.